LOCUS       sequence-436625-        7774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To track the natural NarL promoter's transcriptional dynamics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7774)
  AUTHORS   Dash S, Jagadeesan R, Baptista ISC, Chauhan V, Kandavalli V, 
            Oliveira SMD, Ribeiro AS
  TITLE     A library of reporters of the global regulators of gene expression 
            in Escherichia coli.
  JOURNAL   mSystems. 2024 Jun 18;9(6):e0006524. doi: 10.1128/msystems.00065-24.
            Epub 2024 Apr 30.
  PUBMED    38687030
REFERENCE   2  (bases 1 to 7774)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "mSystems.";
            date: "2024-06-18"; volume: "9(6):e0006524. doi"; pages: " 
            10.1128/msystems.00065-24. Epub 2024 Apr 30"
COMMENT     Sequence Label: NarL-GFP
FEATURES             Location/Qualifiers
     source          1..7774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1119..2090
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     terminator      complement(2248..2334)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2745..2753
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2759..3472
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2789..2817)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3412..3432
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3535..3629
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3764..4423)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4338..4357
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4424..4526)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      5384..5998
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      5475..6006
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      6074..6293
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             6384..7139
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    7142..7392
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             join(7718..7774,1..1119)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
ORIGIN
        1 atgacgaagg ctatcgccat tgcacagttt aatgatgaca gcccggaagc gaggaaaata
       61 acccggcgct ggagaatagg tgaagcagcg gatttagttg gggtttcttc tcaggctatc
      121 agagatgccg agaaagcagg gcgactaccg cacccggata tggaaattcg aggacgggtt
      181 gagcaacgtg ttggttatac aattgaacaa attaatcata tgcgtgatgt gtttggtacg
      241 cgattgcgac gtgctgaaga cgtatttcca ccggtgatcg gggttgctgc ccataaaggt
      301 ggcgtttaca aaacctcagt ttctgttcat cttgctcagg atctggctct gaaggggcta
      361 cgtgttttgc tcgtggaagg taacgacccc cagggaacag cctcaatgta tcacggatgg
      421 gtaccagatc ttcatattca tgcagaagac actctcctgc ctttctatct tggggaaaag
      481 gacgatgtca cttatgcaat aaagcccact tgctggccgg ggcttgacat tattccttcc
      541 tgtctggctc tgcaccgtat tgaaactgag ttaatgggca aatttgatga aggtaaactg
      601 cccaccgatc cacacctgat gctccgactg gccattgaaa ctgttgctca tgactatgat
      661 gtcatagtta ttgacagcgc gcctaacctg ggtatcggca cgattaatgt cgtatgtgct
      721 gctgatgtgc tgattgttcc cacgcctgct gagttgtttg actacacctc cgcactgcag
      781 tttttcgata tgcttcgtga tctgctcaag aacgttgatc ttaaagggtt cgagcctgat
      841 gtacgtattt tgcttaccaa atacagcaat agtaatggct ctcagtcccc gtggatggag
      901 gagcaaattc gggatgcctg gggaagcatg gttctaaaaa atgttgtacg tgaaacggat
      961 gaagttggta aaggtcagat ccggatgaga actgtttttg aacaggccat tgatcaacgc
     1021 tcttcaactg gtgcctggag aaatgctctt tctatttggg aacctgtctg caatgaaatt
     1081 ttcgatcgtc tgattaaacc acgctgggag attagataat gaagcgtgcg cctgttattc
     1141 caaaacatac gctcaatact caaccggttg aagatacttc gttatcgaca ccagctgccc
     1201 cgatggtgga ttcgttaatt gcgcgcgtag gagtaatggc tcgcggtaat gccattactt
     1261 tgcctgtatg tggtcgggat gtgaagttta ctcttgaagt gctccggggt gatagtgttg
     1321 agaagacctc tcgggtatgg tcaggtaatg aacgtgacca ggagctgctt actgaggacg
     1381 cactggatga tctcatccct tcttttctac tgactggtca acagacaccg gcgttcggtc
     1441 gaagagtatc tggtgtcata gaaattgccg atgggagtcg ccgtcgtaaa gctgctgcac
     1501 ttaccgaaag tgattatcgt gttctggttg gcgagctgga tgatgagcag atggctgcat
     1561 tatccagatt gggtaacgat tatcgcccaa caagtgctta tgaacgtggt cagcgttatg
     1621 caagccgatt gcagaatgaa tttgctggaa atatttctgc gctggctgat gcggaaaata
     1681 tttcacgtaa gattattacc cgctgtatca acaccgccaa attgcctaaa tcagttgttg
     1741 ctcttttttc tcaccccggt gaactatctg cccggtcagg tgatgcactt caaaaagcct
     1801 ttacagataa agaggaatta cttaagcagc aggcatctaa ccttcatgag cagaaaaaag
     1861 ctggggtgat atttgaagct gaagaagtta tcactctttt aacttctgtg cttaaaacgt
     1921 catctgcatc aagaactagt ttaagctcac gacatcagtt tgctcctgga gcgacagtat
     1981 tgtataaggg cgataaaatg gtgcttaacc tggacaggtc ccgtgttcca actgagtgta
     2041 tagagaaaat tgaggccatt cttaaggaac ttgaaaagcc agcaccctga tgcgaccacg
     2101 ttttagtcta cgtttatctg tctttactta atgtcctttg ttacaggcca gaaagcataa
     2161 ctggcctgaa tattctctct gggccccact tctttgagtg agctgatacc gctcgccgca
     2221 gccgaacgcc taagtctagg gcggcggatt tgtcctactc aggagagcgt tcaccgacaa
     2281 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcca
     2341 cgcgttggat taatattgat tctctcgaga cggtccctag gagacacctc tgatactcgt
     2401 ttcgatgtct gccaccttag tgtctgtagc taaaggcaat ttgatgtaaa tcaaacgata
     2461 agataacttt atctatcatt gatatttatc attacccata gtgagtacag tgacttcata
     2521 aaaattatga gatttttcac ggtgctgtaa aatccctacc cttaccgatg taaagcgact
     2581 aaccacacgg caaataagga gtaactcttt ccgggtatgg gtatacttca gccaatagcc
     2641 gagaatactg ccattccaga atgtatcgtc acattcatta aggttattgc tcatttaaag
     2701 cctgaaggaa gaggtttaca tgccagttgt agatgacact aaattaagga ggtataatat
     2761 gcgtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     2821 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
     2881 acttaccctt aaatttattt gcactactgg aaaactacct gttccgtggc caacacttgt
     2941 cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcaca tgaaacagca
     3001 tgactttttc aagagtgcca tgcccgaagg ttacgtacag gaaagaacta tatttttcaa
     3061 agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa
     3121 tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt
     3181 ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat
     3241 caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca
     3301 ttatcaacaa aatactccga ttggcgatgg ccctgtcctt ttaccagaca accattacct
     3361 gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct
     3421 tgagtttgta accgctgctg ggattacaca tggcatggat gaactataca aataacagct
     3481 gggtatcgaa ttcaagtgac tgcagaacag tagtacttgc acagacgtcg cttggactcc
     3541 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
     3601 gccgccgggc gttttttatt ggtgagaatc caagcatact cgtcgagacc aattctcatg
     3661 tttgacagct tatcatcgaa tttctgccat tcatccgctt attatcactt attcaggcgt
     3721 agcaaccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc
     3781 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     3841 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     3901 tgcccatcgt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     3961 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
     4021 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     4081 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     4141 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     4201 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     4261 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     4321 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     4381 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     4441 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     4501 agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct
     4561 tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag
     4621 gtatttattc gcgataagct catggagcgg cgtaaccgtc gcacaggaag gacagagaaa
     4681 gcgcggatct gggaagtgac ggacagaacg gtcaggacct ggattgggga ggcggttgcc
     4741 gccgctgctg ctgacggtgt gacgttctct gttccggtca caccacatac gttccgccat
     4801 tcctatgcga tgcacatgct gtatgccggt ataccgctga aagttctgca aagcctgatg
     4861 ggacataagt ccatcagttc aacggaagtc tacacgaagg tttttgcgct ggatgtggct
     4921 gcccggcacc gggtgcagtt tgcgatgccg gagtctgatg cggttgcgat gctgaaacaa
     4981 ttatcctgag aataaatgcc ttggccttta tatggaaatg tggaactgag tggatatgct
     5041 gtttttgtct gttaaacaga gaagctggct gttatccact gagaagcgaa cgaaacagtc
     5101 gggaaaatct cccattatcg tagagatccg cattattaat ctcaggagcc tgtgtagcgt
     5161 ttataggaag tagtgttctg tcatgatgcc agcaagcggt aacgaaaacg atttgaatat
     5221 gccttcagga acaatagaaa tcttcgtgcg gtgttacgtt gaagtggagc ggattatgtc
     5281 agcaatggac agaacaacct aatgaacaca gaaccatgat gtggtctgtc cttttacagc
     5341 cagtagtgct cgccgcagtc gagcgacagg gcgaagccct cggctggttg ccctcgccgc
     5401 tgggctggcg gccgtctatg gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc
     5461 gagacaccgc ggccggccgc cggcgttgtg gatacctcgc ggaaaacttg gccctcactg
     5521 acagatgagg ggcggacgtt gacacttgag gggccgactc acccggcgcg gcgttgacag
     5581 atgaggggca ggctcgattt cggccggcga cgtggagctg gccagcctcg caaatcggcg
     5641 aaaacgcctg attttacgcg agtttcccac agatgatgtg gacaagcctg gggataagtg
     5701 ccctgcggta ttgacacttg aggggcgcga ctactgacag atgaggggcg cgatccttga
     5761 cacttgaggg gcagagtgct gacagatgag gggcgcacct attgacattt gaggggctgt
     5821 ccacaggcag aaaatccagc atttgcaagg gtttccgccc gtttttcggc caccgctaac
     5881 ctgtctttta acctgctttt aaaccaatat ttataaacct tgtttttaac cagggctgcg
     5941 ccctgtgcgc gtgaccgcgc acgccgaagg ggggtgcccc cccttctcga accctcccgg
     6001 tcgagtgagc gaggaagcac cagggaacag cacttatata ttctgcttac acacgatgcc
     6061 tgaaaaaact tcccttgggg ttatccactt atccacgggg atatttttat aattattttt
     6121 tttatagttt ttagatcttc ttttttagag cgccttgtag gcctttatcc atgctggttc
     6181 tagagaaggt gttgtgacaa attgcccttt cagtgtgaca aatcaccctc aaatgacagt
     6241 cctgtctgtg acaaattgcc cttaaccctg tgacaaattg ccctcagaag aagctgtttt
     6301 ttcacaaagt tatccctgct tattgactct tttttattta gtgtgacaat ctaaaaactt
     6361 gtcacacttc acatggatct gtcatggcgg aaacagcggt tatcaatcac aagaaacgta
     6421 aaaatagccc gcgaatcgtc cagtcaaacg acctcactga ggcggcatat agtctctccc
     6481 gggatcaaaa acgtatgctg tatctgttcg ttgaccagat cagaaaatct gatggcaccc
     6541 tacaggaaca tgacggtatc tgcgagatcc atgttgctaa atatgctgaa atattcggat
     6601 tgacctctgc ggaagccagt aaggatatac ggcaggcatt gaagagtttc gcggggaagg
     6661 aagtggtttt ttatcgccct gaagaggatg ccggcgatga aaaaggctat gaatcttttc
     6721 cttggtttat caaacgtgcg cacagtccat ccagagggct ttacagtgta catatcaacc
     6781 catatctcat tcccttcttt atcgggttac agaaccggtt tacgcagttt cggcttagtg
     6841 aaacaaaaga aatcaccaat ccgtatgcca tgcgtttata cgaatccctg tgtcagtatc
     6901 gtaagccgga tggctcaggc atcgtctctc tgaaaatcga ctggatcata gagcgttacc
     6961 agctgcctca aagttaccag cgtatgcctg acttccgccg ccgcttcctg caggtctgtg
     7021 ttaatgagat caacagcaga actccaatgc gcctctcata cattgagaaa aagaaaggcc
     7081 gccagacgac tcatatcgta ttttccttcc gcgatatcac ttccatgacg acaggatagt
     7141 ctgagggtta tctgtcacag atttgagggt ggttcgtcac atttgttctg acctactgag
     7201 ggtaatttgt cacagttttg ctgtttcctt cagcctgcat ggattttctc atactttttg
     7261 aactgtaatt tttaaggaag ccaaatttga gggcagtttg tcacagttga tttccttctc
     7321 tttcccttcg tcatgtgact tgatatcggg ggttagttcg tcatcattga tgagggttga
     7381 ttatcacagt ttattactct gaattggcta tccgcgtgtg tacctctacc tggagttttt
     7441 cccacggtgg atatttcttc ttgcgctgag cgtaagagct atctgacaga acagttcttc
     7501 tttgcttcct cgccagttcg ctcgctatgc tcggttacac ggctgcggcg agcgctagtg
     7561 ataataagtg actgaggtat gtgctcttct tatctccttt tgtagtgttg ctcttatttt
     7621 aaacaacttt gcggtttttt gatgactttg cgattttgtt gttgctttgc agtaaattgc
     7681 aagatttaat aaaaaaacgc aaagcaatga ttaaaggatg ttcagaatga aactcatgga
     7741 aacacttaac cagtgcataa acgctggtca tgaa
//
